CHEMOPROTECTIVE ROLE OF BOVINE LACTOFERRICIN AGAINST 7, 12 DIMETHYLBENZ (A) ANTHRACENE INDUCED SKIN CANCER IN FEMALE SWISS ALBINO MICE
Keywords:
Chemoprevention, Lfcin B, Antioxidants, Liver enzymes, Kidney function markers, GlucoseAbstract
Objective: To investigate the chemo-preventive effect of subcutaneous injection of bovine lactofericin (LfcinB) against 7, 12 dimethylbenz (a) anthracene (DMBA) induced carcinogenesis in swiss albino female mice.
Methods: 30 animals were divided into 3 groups. 200μl of DMBA (0.025μg/μl) per animal three times per week as topical was used to induce skin cancer in both groups II and III. In addition to DMBA, group III was protected with subcutaneous injection of LfcinB (20μg/μl) while group I served as control. At the end of 16th week lysates/blood/serum were subjected to various tests such as antioxidant, glucose, liver enzymes and kidney marker parameters.
Results: Topical application of 200μl DMBA (0.025μg/μl) per animal three times per week has produced 100% tumor incidence in DMBA treated group at the end of 16th week whereas only 20% developed tumors in the LfcinB+DMBA treated group. Antioxidants level were found to be significantly (P<0.01) depleted in reduced glutathione (GSH), glutathione reductase (GR), glutathione peroxidase (GPx), catalase (CAT) and liver enzymes, Alaninine aminotransferase (ALT) and Alkaline phosphatase (ALP). Subcutaneous administration of 200μl of LfcinB (20μg/μl) significantly (P<0.01) normalized the antioxidants (GSH, GR, GPx and CAT) levels.
Conclusion: LfcinB exerts its chemoprotective effect through acting as an antioxidant, thereby inhibiting carcinogenesis, hepatocellular and renal damage. Lfcin B may act as a promising chemoprotective agent against DMBA induced skin cancer.
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F’guyer S, Afaq F, Mukhtar H. Photochemoprevention of skin cancer by botanical agents. Phtodermatol Photoimmunol Photomed 2003;19:56-72.
Dorai T, Aggarwal BB. Role of chemopreventive agents in cancer therapy. Cancer Lett 2004;215:129-40.
Crowell JA. The chemopreventive agent development research program in the division of cancer prevention of the US National Cancer Institute: an overview. Eur J Cancer 2005;41:1889-910.
Steward WP, Brown K. Cancer chemoprevention: a rapidly evolving field. Br J Cancer 2013;109:1-7.
Yu S, Kong AN. Targeting carcinogen metabolism by dietary cancer preventive compounds. Curr Cancer Drug Targets 2007;7:416-24.
Hauser AT, Jung M. Targeting epigenetic mechanisms: the potential of natural products in cancer chemoprevention. Planta Med 2008;74:1593-601.
Bellamy W, Takase M, Yamauchi K, Wakabayashi H, Kawase K, Tomita M. Identification of the bactericidal domain of lactoferrin. Biochim Biophys Acta 1992;1121:130-6.
Kuwata H, Yip TT, Tomita M, Hutchens TW. Direct evidence of the generation in the human stomach of an antimicrobial peptide domain (lactoferricin) from ingested lactoferrin. Biochem Biophys Acta 1998;1429:129-41.
Tomita M, Bellamy W, Takase M, Yamauchi K, Wakabayashi H, Kawase K. Potent antibacterial peptides generated by pepsin digestion of bovine lactoferrin. J Dairy Sci 1991;74:4137-42.
Bellamy W, Takase M, Wakabayashi H, Kawase K, Tomita M. Antibacterial spectrum of lactoferricin B, a potent bactericidal peptide derived from the N-terminal region of bovine lactoferrin. J Appl Bacteriol 1992;73:472-9.
Yamauchi K, Tomita M, Giehl TJ, Ellison RT. Antibacterial activity of lactoferrin and a pepsin-derived lactoferrin peptide fragment. Infect Immun 1993;61:719-28.
Yoo YC, Watanabe S, Watanabe R, Hata K, Shimazaki K, Azuma I. Bovine lactoferrin and lactoferricin, a peptide derived from bovine lactoferrin, inhibit tumor metastasis in mice. Jpn J Cancer Res 1997;88:184-90.
Cho E, Smith-Warner SA, Spiegelman D, Beeson WL, Van den Brandt PA, Colditz GA, et al. Dairy foods, calcium, and colorectal cancer: a pooled analysis of 10 cohort studies. J Natl Cancer Inst 2004;96:1015-22.
Tsuda H, Sekine K, Nakamura J, Ushida Y, Kuhara T, Takasuka N, et al. Inhibition of azoxymethane-initiated colon tumor and aberrant crypt foci development by bovine lactoferrin administration in F344 rats. Adv Exp Med Biol 1998;443:273-84.
Perdigon G, de Moreno de LeBlanc A, Valdez J, Rachid M. Role of yogurt in the prevention of colon cancer. Eur J Clin Nutr 2002;56 Suppl 3:65-8.
Krueger H, McLean D, Williams D. Skin cancers. Prog Exp Tumor Res 2008;40:111-21.
Boukamp P. Non-melanoma skin cancer. What drives tumor development and progression? Carcinogenesis 2005;26:1657-67.
Greenlee RT, Hill-Harmon MB, Murray T, Thun M. Cancer statistics. CA Cancer J Clin 2001;51:15-36.
Stern RS. Prevalence of a history of skin cancer in 2007: results of an incidence-based model. Arch Dermatol 2010;146:279-82.
Kalicharran S, Diab RD, Sokolic F. Trends in total ozone over southern African stations between 1979 and 1991. Geophys Res Lett 1993;20:2877-80.
Staples MP, Elwood M, Burton RC, Williams JL, Marks R, Giles GG. Non-melanoma skin cancer in Australia: the 2002 national survey and trends since 1985. Med J Aust 2006;184:6-10.
Deo SV, Hazarika S, Shukla NK, Kumar S, Kar M, Samaiya A. Surgical management of skin cancers: experience from a regional cancer center in North India. Indian J Cancer 2005;42:145-50.
Miyata M, Furukawa M, Takahashi K, Gonzalez FJ, Yamazoe Y. Mechanism of 7, 12-dimethylbenz[a] anthracene induced immunotoxicity: Role of metabolic activation at the target organ. Jpn J Pharmacol 2001;86:302-9.
Das RK, Bhattacharya S. Inhibition of DMBA-croton oil two-stage mouse skin carcinogenesis by diphenyl methyl selenocyanate through modulation of cutaneous oxidative stress and inhibition of nitric oxide production. Asian Pacific J Cancer Prevention 2004;5:151-8.
Nigam N, Prasad S, Shukla Y. Preventive effects of lupeol on DMBA induced DNA alkylation damage in mouse skin. Food Chem Toxicol 2007;45:2331-5.
Rastogi S, Shukla Y, Paul BN, Chowdhuri DK, Khanna SK, Das M. Protective effect of Ocimum sanctum on 3-methylcholanthrene, 7, 12-dimethylbenz (a) anthracene and aflatoxin B1 induced skin tumorigenesis in mice. Toxicol Appl Pharmacol 2007;224:228-40.
Barrera G. Oxidative stress and lipid peroxidation products in cancer progression and therapy. ISRN Oncol 2012:1-21. Doi:10.5402/2012/137289. [Article in Press]
Lu W, Ogasawara MA, Huang P. Models of reactive oxygen species in cancer. Drug Discovery Today Dis Models 2007;4:67-73.
Ishii N. Role of oxidative stress from mitochondria on aging and cancer. Cornea 2007;26(9, Suppl 1):3-9.
Khan AR, Taneja P. Cationic peptide lactoferricin B inhibits glutathione s-transferase p1 from human placenta and breast cancer cell line MDA-MB-231 preventing anticancer drug metabolism. Int J Pharm Pharm Sci 2015;7:238-41.
Ellman GL. Tissue sulfhydryl groups. Arch Biochem Biophys 1959;82:70-7.
Habig WH, Pabst MJ, Jakoby WB. Glutathione-stransferases: the first enzymatic step in mercapturic acid formation. J Biol Chem 1974;249:7130-9.
Takahara S, Hamilton BH, Nell JV, Ogubra TY, Nishimura ET. Hypocatalasemia: a new genetic carrier state. J Clin Invest 1960;39:610-9.
Misra HP, Fridovich I. The role of superoxide anion in the autoxidation of epinephrine and a simple assay for superoxide dismutase. J Biol Chem 1972;247:3170-5.
Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxidation in animal tissue by thiobarbituric acid reaction. Anal Biochem 1979;95:351-8.
Rotruck JT, Pope AL, Gauther HE, Swanson AB, Hafeman DG, Hoekstna WG. Selenium: biochemical roles as a component of glutathione peroxidase. Science 1973;179:588-90.
Carlberg I, Mannervik B. Purification and characterization of the flavoenzyme glutathione reductase from rat liver. J Biol Chem 1975;250:5475-80.
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Estimation of protein by using Folin phenol reagent. J Biol Chem 1951;193:267-75.
Trinder P. Determination of glucose in blood using glucose oxidase with an alternative oxygen receptor. Ann Clin Biochem 1969;6:24-7.
Schumann G, Bonora R, Ceriotti F, Férard G, Ferrero CA, Franck PF. IFCC primary reference procedures for the measurement of catalytic activity concentrations of enzymes at 37 °C Part 5. Reference procedure for the measurement of catalytic concentration of aspartate aminotransferase. Clin Chem Lab Med 2002;40:725-33.
Schumann G, Bonora R, Ceriotti F, Férard G, Ferrero CA, Franck PF, et al. IFCC primary reference procedures for the measurement of catalytic activity concentrations of enzymes at 37 °C. Part 4. Reference procedure for the measurement of catalytic concentration of alanine aminotransferase. Clin Chem Lab Med 2002;40:718-24.
Tietz NW, Burtis CA, Duncan P, Ervin K, Petitclerc CJ, Rinker AD, et al. A reference method for measurement of alkaline phosphatase activity in human serum. Clin Chem 1983;29:51-76.
Sporn MB, Suh N. Chemoprevention of cancer. Carcinogenesis 2000;21:525-30.
Abel EL, Angel JM, Kiguchi K, DiGiovanni J. Multi-stage chemical carcinogenesis in mouse skin: fundamentals and applications. Nat Protoc 2009;4:1350-62.
Lushchak VI. Glutathione homeostasis and functions: potential targets for medical interventions. J Amino Acids 2012;736837:1-26.
Singh S, Khan AR, Gupta AK. Role of glutathione in cancer pathophysiology and therapeutic interventions. J Exp Ther Oncol 2012;9:303-16.
Anderson ME. Glutathione: an overview of biosynthesis and modulation. Chem Biol Interact 1998;111-112:1-14.
Mullineaux PM, Creissen G. Glutathione reductase: regulation and role in oxidative stress. In: Oxidative Stress and the Molecular Biology of Antioxidant Defenses. Cold Spring Harbor Laboratory Press: Cold Spring Harbor, New York; 1997. p. 667-713.
Townsend DM, Tew KD, Tapiero H. The importance of glutathione in human disease. Biomed Pharmacother 2003;57:145-55.
Oberley TD. Oxidative damage and cancer. Am J Pathol 2002;160:403-8.
Perchellet JP, Perchellet M. Antioxidant and multistage carcinogenesis in mouse skin. Free Radical Biol Med 1989;7:377-408.
Elangovan V, Balasubramanian S, Sekar N, Govindasamy S. Studies on the chemopreventive potential of some naturally occurring bioflavonoids in 7,12-dimethylbenz(a)anthracene induced carcinogenesis in mouse skin. J Clin Biochem Nutr 1994;17:153-60.
Halliwel B. Oxidative stress and cancer: have we moved forward. Biochem J 2007;401:1-11.
Niitsu Y, Takahashi Y, Saito T, Hirata Y, Arisato N, Maruyama H, et al. Serum glutathione-S-transferase-pi as a tumor marker for gastrointestinal malignancies. Cancer 1989;63:317-23.
Randall BJ, Angus B, Akiba R, Hall A, Cattan AR, Proctor SJ, et al. Glutathione S-transferase (placental) as a marker of transformation in the human cervix uteri: an immunohistochemical study. Br J Cancer 1990;62:614-8.
Kantor RR, Giardina SL, Bartolazzi A, Townsend AJ, Myers CE, Cowan KH, et al. Monoclonal antibodies to glutathione S-transferase pi-immunohistochemical analysis of human tissues and cancers. Int J Cancer 1991;47:193-201.
Santa T, Isobe K, Yamauchi M, Nakashima I, Takagi. Expression of MDR1 and glutathione S-transferase-pi genes and chemosensitivities in human gastrointestinal cancer. Cancer 1992;69:941-6.
Green JA, Robertson LJ, Clark AH. Glutathione S-transferase expression in benign and malignant ovarian tumors. Br J Cancer 1993;68:235-9.
Inoue T, Ishida T, Sugio K, Maehara Y, Sugimachi K. Glutathione S-transferase Pi is a powerful indicator in chemotherapy of human lung squamous cell carcinoma. Respiration 1995;62:223-37.
Bentz BG, Haines GK, Radosevich JA. Glutathione S-transferase pi in squamous cell carcinoma of the head and neck. Laryngoscope 2000;110:1642-7.
Trachte AL, Suthers SE, Lerner MR, Hanas JS, Jupe ER, Sienko AE, et al. Increased expression of alpha-1-antitrypsin, glutathione S-transferase pi and vascular endothelial growth factor in human pancreatic adenocarcinoma. Am J Surg 2002;184:647-8.
Mimic T, Mimic-Oka J, Savic-Radojevic A, Opacic M, Pljesa M, Dragicevic D, et al. Glutathione S-transferase T1-1 activity upregulated in transitional cell carcinoma of urinary bladder. Urology 2005;65:1035-40.
Arai T, Miyoshi Y, Kim SJ, Taguchi T, Tamaki Y, Noguchi S. Association of GSTP1 CpG islands hypermethylation with poor prognosis in human breast cancers. Breast Cancer Res Treat 2006;100:169-76.
Raha S, Robinson BH. Mitochondria, oxygen, free radical, diseases and aging. Trends Biochem Sci 2000;25:502-8.
James LP, Mayeux PR, Hinson JA. Acetaminophen-induced hepatotoxicity. Drug Metab Dispos 2003;31:1499-506.
Fakurazi S, Hairuszah I, Nanthini U. Moringa oleifera Lam prevents acetaminophen induced liver injury through restoration of glutathione level. Food Chem Toxicol 2008;46:2611-5.
Okuda K. Hepatocellular carcinoma: clinicopathological aspects. J Gastroenterol Hepatol 1997;(9, Suppl 2):314-8.
Shyamal S, Latha PG, Shine VJ, Suja SR, Rajasekharan S, Ganga Devi T. Hepatoprotective effects of Pittosporum neelgherrense Wightand Arn.,a popular Indian ethnomedicine. J Ethnopharmacol 2006;107:151-5.
Panteghini M. Aspartate amino transferaseisoenzymes. Clin Biochem 1990;23:311-9.